CN218026002U

CN218026002U - Cell sorting device

Info

Publication number: CN218026002U
Application number: CN202221782866.9U
Authority: CN
Inventors: 胡璧梁; 尹果; 谭亮; 翁宽; 肖泽玺; 江宏伟
Original assignee: Chutian Siweikang Gene Technology Changsha Co ltd
Current assignee: Changsha Chusi Weikang Intelligent Technology Co.,Ltd.
Priority date: 2022-07-08
Filing date: 2022-07-08
Publication date: 2022-12-13
Anticipated expiration: 2032-07-08

Abstract

The utility model discloses a cell sorting device, including control box, wabbler mechanism, be arranged in carrying the cell liquid to select separately the pump in the container, wabbler mechanism is equipped with the arc that is used for bearing cell liquid including the rocking frame that is used for placing the separation container and the drive assembly that sways that is used for driving the rocking frame, selects separately the container, is equipped with the magnetism that is used for changing separation container magnetic field intensity on the rocking frame and inhales the mechanism, inhales the arc wall that is equipped with the arc looks adaptation with the separation container in the mechanism. The cell sorting device is convenient to operate, and can accelerate the combination speed of magnetic beads and cells and the sorting speed.

Description

Cell sorting device

Technical Field

The utility model relates to a food, pharmaceutical packaging mechanical equipment technical field especially relate to a cell sorting unit.

Background

Cell sorting is the separation of a cell from a multicellular sample. Cells are usually labeled with antibodies, while the antibodies are fluorescently labeled or linked to immunomagnetic beads or others, using the properties of fluorescence and immunomagnetic beads to separate cells.

At present, common immunomagnetic bead cell sorting equipment at home and abroad adopts a first mode, flow cell sorting separates luminescent particle subgroups according to the fluorescence intensity and wavelength of emitted light and can realize monoclonal sorting, the sorting speed is low, instrument parameters need to be adjusted before use, the operation is complex, and the technical difficulty is high; the method II is characterized in that the nano magnetic beads are combined with cells, the combination speed is low, no demagnetized bead process is needed after separation, and when the separation column filled with the beads is passed, the separation column is easy to block, and the separation speed is low; and in the third mode, the magnetic beads are adsorbed by the flat-plate magnet, the equipment is manually operated, so that manual intervention is more, errors are easy to occur, and data recording and tracking cannot be completed.

The application will explain a cell sorting device, which adopts an immunomagnetic bead cell sorting method, and combines 2-8 μm immunomagnetic beads with cells to make them adsorbed when passing through a stable high gradient magnetic field, thereby realizing cell sorting. The device has the advantages of simple operation, high sorting speed and good cell activity, and can separate high-purity, high-activity and high-yield cells from complex cell mixtures within a few minutes. The cell sorting equipment can solve the problems of low sorting speed, complex operation, much manual intervention, no data recording in the process and the like of various cell sorting equipment in the current market.

SUMMERY OF THE UTILITY MODEL

The to-be-solved technical problem of the utility model is to overcome the not enough of prior art, provide a convenient operation, can accelerate magnetic bead and cell combination speed and can accelerate the cell sorting unit of sorting speed.

In order to solve the technical problem, the utility model adopts the following technical scheme:

the utility model provides a cell sorting unit, includes control box, wabbler mechanism, is arranged in carrying the cell liquid to select separately the pump in the container, wabbler mechanism is including the rocking frame that is used for placing the separation container and the drive assembly that sways that is used for driving the rocking frame to sway, the separation container is equipped with the arc portion that is used for bearing cell liquid, be equipped with the magnetism that is used for changing separation container magnetic field intensity on the rocking frame and inhale the mechanism, inhale the arc wall that is equipped with the arc looks adaptation with the separation container in the mechanism.

As a further improvement of the technical scheme:

the magnetic attraction mechanism comprises a lifting seat arranged on the swing frame, a magnet assembly arranged on the lifting seat and a lifting driving assembly used for driving the lifting seat to lift, the magnet assembly is located below the clamping groove, and the arc-shaped groove is formed in the upper surface of the magnet assembly.

The lifting driving assembly comprises a lifting driving motor, a gear and a rack, the lifting driving motor is installed on the swing frame, the gear is connected with an output shaft of the lifting driving motor, and the rack is arranged on the lifting seat and meshed with the gear.

The swing frame is provided with a guide groove, the lifting seat is provided with a rolling piece, the rolling piece is in contact with the inner wall of the guide groove, and two ends of the guide groove are respectively provided with a limiting piece.

The control box is provided with a digital display module which is electrically connected with each other and a code scanning gun which is used for scanning the code liquid containing bag and the collecting bag.

Two clamping grooves are formed in the swing frame and arranged in parallel.

Cell sorting unit still includes trunk line, two first minute pipelines, two flourishing liquid bags, two sorting containers, two collection bags, two second minute pipelines, two third minute pipelines and two fourth minute pipelines, flourishing liquid bag is hung and is located on the stores pylon, collect the bag and place on placing the bench, two sorting containers blocks respectively in locating two draw-in grooves on the rocking frame, two flourishing liquid bag is connected with the inlet end of pump, two through two first minute pipelines respectively sorting containers's inlet end is connected with the other end of the outlet end of pump respectively, and the liquid outlet end is connected with the one end of trunk line through two third minute pipelines respectively, two collecting bags is connected with the other end of trunk line through two fourth minute pipelines respectively, each all be equipped with flow control valve on the minute pipeline.

And a magnetic bead injection port and a breathing air-permeable port are arranged on the separation container.

Compared with the prior art, the utility model has the advantages of:

the cell sorting device of the utility model is produced by arranging the liquid containing bag and the collecting bag on the control box, placing the sorting container on the swing frame, and connecting the liquid containing bag, the pump, the sorting container and the collecting bag through pipelines; and injecting magnetic beads into the sorting container; in the cell sorting process, the sorting container is rocked through the rocking mechanism, so that the combination speed of the magnetic beads and the cells can be increased; after the magnetic beads are combined with the cells, the magnetic attraction mechanism applies larger magnetic field intensity to the sorting container, the magnetic beads and the cell combination in the sorting container are adsorbed on one side of the sorting container close to the magnetic attraction mechanism, then the sorting container is inclined through the swing frame, and the liquid in the sorting container is poured out, namely the combination of the magnetic beads and the cells is left; then, the cells in the combination of the magnetic beads and the cells are sorted out by repeatedly injecting a buffer solution into the sorting container, changing the magnetic field strength of the sorting container by the magnetic attraction mechanism, and pouring out the supernatant. The cell sorting device is convenient to operate, and can accelerate the combination speed of magnetic beads and cells and the sorting speed.

Drawings

FIG. 1 is a schematic perspective view of the cell sorting device of the present invention.

FIG. 2 is a schematic front view of the cell sorting device according to the present invention.

FIG. 3 is a schematic diagram of the side view structure of the cell sorting device of the present invention.

FIG. 4 is a schematic top view of the cell sorting apparatus according to the present invention.

FIG. 5 is a schematic sectional view of the rocking arm of the cell sorting apparatus according to the present invention (the magnet assembly is close to the sorting container).

FIG. 6 is a schematic sectional view of the rocking arm of the cell sorting apparatus according to the present invention (the magnet assembly is far from the sorting container).

FIG. 7 is a schematic view showing a first view angle structure inside a rocking frame of the cell sorting apparatus according to the present invention.

FIG. 8 is a schematic view showing a second view angle inside the rocking frame of the cell sorting apparatus according to the present invention.

FIG. 9 is a schematic view showing the structure of a sorting container in a rocking frame of the cell sorting apparatus according to the present invention.

The reference numerals in the figures denote:

1. a control box; 11. a pump; 12. a flow regulating valve; 13. a hanger; 14. a placing table; 15. a digital display module; 16. a code scanning gun; 2. a main pipeline; 3. a liquid containing bag; 31. a first branch duct; 4. a sorting container; 41. a magnetic bead injection port; 42. a breathing vent; 43. an arcuate portion; 5. a collection bag; 51. a fourth branch pipe; 6. a second branch pipe; 7. a third branch pipeline; 8. a rocking mechanism; 81. a swing frame; 811. a card slot; 812. a guide groove; 813. a limiting member; 82. a swing drive assembly; 9. a magnetic attraction mechanism; 91. a lifting seat; 911. a rolling member; 92. a magnet assembly; 921. an arc-shaped slot; 93. a lift drive assembly; 931. a lifting drive motor; 932. a gear; 933. a rack.

Detailed Description

The invention will be described in further detail with reference to the drawings and specific examples.

As used in this disclosure and in the claims, the terms "a," "an," "the," and/or "the" are not intended to be inclusive in the singular, but rather are inclusive in the plural unless the context clearly dictates otherwise. The use of "first," "second," and similar terms in this disclosure is not intended to indicate any order, quantity, or importance, but rather is used to distinguish one element from another. Likewise, the word "comprising" or "comprises", and the like, means that the element or item listed before the word covers the element or item listed after the word and its equivalents, but does not exclude other elements or items. The terms "connected" or "coupled" and the like are not restricted to physical or mechanical connections, but may include electrical connections, whether direct or indirect.

The first embodiment is as follows:

fig. 1 to 9 show an embodiment of the cell sorting device of the present invention, this cell sorting device includes control box 1, wabbler mechanism 8, a pump 11 for carrying the cell liquid into the sorting container 4, wabbler mechanism 8 includes the driving component 82 that sways that is used for placing the swaying frame 81 of sorting container 4 and is used for driving swaying frame 81 to sway, sorting container 4 is equipped with the arc 43 that is used for bearing cell liquid, be equipped with the magnetism mechanism 9 of inhaling that is used for changing 4 magnetic field intensity of sorting container on the swaying frame 81, it is equipped with the arc 921 with the arc 43 looks adaptation of sorting container on the magnetism mechanism 9.

The sorting container 4 is provided with an arc part 43 for supporting the cell sap, the arc part can be designed into a round, oval or other smooth curved surface, and the magnetic attraction mechanism 9 is provided with an arc groove matched with the arc part of the sorting container, so that the magnetic field adsorption force borne by the magnetic beads is more uniform; under the condition of equal perimeter, the area of the arc part is larger than that of the square part, so that the magnetic field adsorption area is larger, and the magnetic bead adsorption speed is higher; the design of the arc-shaped part reduces dead angles, and the impact on the cell liquid in the swing process is smaller.

During production, the liquid containing bag 3 and the collecting bag 5 are arranged on the control box 1, the sorting container 4 is placed on the swing frame 81, and the liquid containing bag 3, the pump 11, the sorting container 4 and the collecting bag 5 are connected through pipelines; and magnetic beads are injected into the sorting container 4; in the cell sorting process, the sorting container 4 is swung by the swinging mechanism 8, so that the combination speed of magnetic beads and cells can be increased; after the magnetic beads are combined with the cells, the magnetic attraction mechanism 9 applies a larger magnetic field strength to the sorting container 4, the magnetic beads and cell combinations in the sorting container 4 are adsorbed on one side of the sorting container 4 close to the magnetic attraction mechanism 9, then the sorting container 4 is inclined through the swing frame 81, the liquid in the sorting container 4 is poured out, and the magnetic beads and cell combinations are left; then, the cells in the bound body of the magnetic beads and the cells are sorted out by repeatedly injecting a buffer solution into the sorting container 4, changing the magnetic field strength of the sorting container 4 by the magnetic attracting mechanism 9, and pouring out the supernatant. The cell sorting device is convenient to operate, and can accelerate the combination speed of magnetic beads and cells and the sorting speed.

The cell sorting device adopts an immunomagnetic bead cell sorting method, and the immunomagnetic beads with the particle size of 2-8 mu m are combined with cells to be adsorbed when passing through a stable high gradient magnetic field, thereby realizing cell sorting. The device has simple operation, high sorting speed and good cell activity, and can separate high-purity, high-activity and high-yield cells from complex cell mixtures within a few minutes. The device can solve the problems of low sorting speed, complex operation, multiple manual interventions, no data recording in the process and the like of various cell sorting devices in the current market.

In this embodiment, as shown in fig. 5 to 8, a clamping groove 811 adapted to the sorting container 4 is disposed on the swing mechanism 8, the magnetic attraction mechanism 9 includes a lifting seat 91 disposed on the swing frame 81, a magnet assembly 92 disposed on the lifting seat 91, and a lifting driving assembly 93 for driving the lifting seat 91 to lift, the magnet assembly 92 is disposed below the clamping groove 811, and an arc-shaped groove 921 is formed on an upper surface of the magnet assembly 92. The clamping groove 811 is matched with the sorting container 4, so that the sorting container 4 can be conveniently taken and placed. The lifting driving component 93 drives the lifting seat 91 to lift so as to drive the magnet component 92 to lift, so that the magnet component 92 moves close to and away from the clamping groove 811, the magnetic field intensity of the sorting container 4 in the clamping groove 811 is changed, and the magnetic beads in the sorting container 4 are attracted and released. Simple structure and convenient operation.

In this embodiment, the lifting driving assembly 93 includes a lifting driving motor 931, a gear 932 and a rack 933, the lifting driving motor 931 is installed on the rocking frame 81, the gear 932 is connected with an output shaft of the lifting driving motor 931, and the rack 933 is arranged on the lifting base 91 and is meshed with the gear 932. The elevating drive motor 931 drives the gear 932 to rotate, and the gear 932 drives the elevating base 91 to move up and down through the meshing transmission effect with the rack 933, so as to realize the elevating movement of the magnet assembly 92.

In this embodiment, as shown in fig. 7 and 8, a guide groove 812 is provided on the swing frame 81, a rolling member 911 is provided on the elevating base 91, and the rolling member 911 contacts with an inner wall of the guide groove 812. The guide groove 812 plays a role in guiding the lifting of the lifting seat 91, and the lifting seat 91 is in contact with the inner wall of the guide groove 812 through the rolling piece 911, so that the abrasion is reduced.

In this embodiment, the two ends of the guide groove 812 are respectively provided with a limiting member 813. The limiting member 813 is used to limit the lifting range of the lifting base 91.

In this embodiment, as shown in fig. 1, the control box 1 is provided with a digital display module 15 and a code scanning gun 16 for scanning the code liquid containing bag 3 and the collecting bag 5, which are electrically connected with each other. Sweep a yard rifle 16 and be used for sweeping a yard flourishing liquid bag 3 and collection bag 5 to the record serial number, with the combination of digital display module 15, the digital management of being convenient for. A wireless or wired code scanning gun 16 may be used; before sorting, the disposable pipeline, the primary sorting container 4, the immunomagnetic beads, the centrifuged leukocyte collection bag 5, the target cell collection bag 5, the waste liquid collection bag 5 and the like are required to be subjected to code scanning and numbering recording; sweep a yard rifle 16 and can be with data transmission to digital display module 15, record, storage key parameter data to guarantee data traceability.

One, two or more engaging grooves 811 are provided on the rocking arm 81 and arranged in parallel. In this embodiment, two locking grooves 811 are preferably provided in the rocking frame 81, and the two locking grooves 811 are arranged in parallel. Two separation container 4 can be placed to two draw-in grooves 811, improves production efficiency. Specifically, two engaging grooves 811 are respectively provided on the bottom surface of the rocking frame 81, and two magnet assemblies 92 are provided on the lifting base 91, and are respectively disposed corresponding to the two engaging grooves 811, that is, one magnet assembly 92 is disposed below each engaging groove 811.

Example two:

as shown in fig. 1 to 4, a cell sorting device, including all the structures of the first embodiment, further including a main pipe 2, two first branch pipes 31, two liquid containing bags 3, two sorting containers 4, two collecting bags 5, two second branch pipes 6, two third branch pipes 7 and two fourth branch pipes 51, the liquid containing bags 3 are hung on a hanging frame 13, the collecting bags 5 are placed on a placing table 14, the two sorting containers 4 are respectively clamped in two clamping grooves 811 on a swinging frame 81, the two liquid containing bags 3 are respectively connected with the inlet end of a pump 11 through the two first branch pipes 31, the liquid inlet ends of the two sorting containers 4 are respectively connected with the other end of the outlet end of the pump 11 through the two second branch pipes 6, the liquid outlet ends are respectively connected with one end of the main pipe 2 through the two third branch pipes 7, the two collecting bags 5 are respectively connected with the other end of the main pipe 2 through the two fourth branch pipes 51, and each branch pipe is provided with a flow regulating valve 12.

The magnet assembly 92 is composed of two semicircular magnets, the outer surface of which is wrapped by a semicircular stainless steel plate or other materials capable of isolating magnetic fields, and the two semicircular magnets are connected together through a connecting plate, so that when the rack 933 moves up and down, the two semicircular magnets can be ensured to rise and fall simultaneously, and the magnetic fields can be provided and removed for the separation container 4.

The cell sorting device comprises the following cell sorting steps: step one, adding cell sap: opening the corresponding flow regulating valve 12 to add cell fluid and optionally buffer solution to one or both of the sorting containers 4; and step two, swinging the sorting container 4: the swing mechanism 8 swings the sorting container 4 for a predetermined time period to combine the cells and the magnetic beads in the sorting container 4; step three, discharging supernatant: the magnetic attracting mechanism 9 attracts the magnetic beads in the sorting container 4; the swing mechanism 8 tilts the sorting container 4 to a predetermined angle; opening the corresponding flow regulating valve 12 to discharge the supernatant in the sorting container 4 into a collecting bag 5; fourthly, resuspending: repeating the second step to resuspend the cells and the bead conjugate; and a fifth step of discharging the cell and magnetic bead combination: the magnetic attraction mechanism 9 releases the magnetic beads in the sorting container 4; the swing mechanism 8 tilts the sorting container 4 to a predetermined angle; the corresponding flow control valve 12 is opened to discharge the cell and magnetic bead conjugates in the sorting container 4 to another collection bag 5.

In this embodiment, the sorting container 4 is provided with a magnetic bead injection port 41 and a breathing gas vent port 42.

The sorting container 4 is made of biocompatible transparent thin-wall plastic through mold opening, and the separating effect on a magnetic field is small due to the transparent thin-wall plastic material. A liquid inlet of the separation container 4 is connected with a hose and a luer connector, is positioned in the middle position of the end part of the separation container 4 and is used for adding cell sap, buffer solution and the like; the liquid outlet of the sorting container 4 is connected with a hose and a luer connector, is positioned at the lower position of the other opposite end part of the sorting container 4 and is used for discharging target cell sap and waste liquid. The breathing vent 42 is connected to a hose and a gas filter and is located at the upper part of the separation container 4 for equalizing the pressure inside the separation container 4 during filling. The magnetic bead injection port 41 is connected to a hose and a luer connector, is located at the upper part of the sorting container 4, and is used for adding magnetic beads into the sorting container 4 before sorting.

Example three:

a cell sorting method is carried out by adopting the cell sorting device in the second embodiment, two liquid containing bags 3 are respectively filled with cell liquid and buffer liquid, magnetic beads are arranged in a sorting container 4, and the cell sorting method comprises the following steps:

s1, adding cell sap: opening the corresponding flow regulating valve 12 to add cell fluid and optionally buffer solution to one or both of the sorting containers 4;

s2, swinging the sorting container 4: the swing mechanism 8 swings the sorting container 4 for a predetermined length of time to combine the cells and the magnetic beads in the sorting container 4;

s3, discharging supernatant: the magnetic attraction mechanism 9 attracts the magnetic beads in the sorting container 4; the swing mechanism 8 tilts the sorting container 4 to a predetermined angle; opening the corresponding flow regulating valve 12 to discharge the supernatant in the sorting container 4 into a collecting bag 5;

s4, resuspending: repeating S2 to resuspend the cells and the magnetic bead conjugate;

s5, expelling cell and magnetic bead conjugates (target): the magnetic attraction mechanism 9 releases the magnetic beads in the sorting container 4; the swing mechanism 8 tilts the sorting container 4 to a predetermined angle; the corresponding flow control valve 12 is opened to discharge the cell and magnetic bead conjugates in the sorting container 4 to another collection bag 5.

The cell sorting method is convenient to operate, and can accelerate the combination speed of magnetic beads and cells and the sorting speed.

An immunomagnetic bead cell sorting method is adopted, and 2-8 mu m immunomagnetic beads are combined with cells and are adsorbed when passing through a stable high gradient magnetic field, so that the cell sorting is realized. The device has the advantages of simple operation, high sorting speed and good cell activity, and can separate high-purity, high-activity and high-yield cells from complex cell mixtures within a few minutes. The cell sorting equipment can solve the problems of low sorting speed, complex operation, much manual intervention, no data recording in the process and the like of various cell sorting equipment in the current market.

White blood cell sorting after centrifugation:

1) Suspending the liquid containing bag 3 (hereinafter referred to as cell liquid bag) containing centrifuged white blood cells and the liquid containing bag 3 (hereinafter referred to as buffer liquid bag) containing buffer solution on the hanging frame 13 at the top of the swing frame 81, and connecting the liquid containing bag 3 and the buffer liquid bag with corresponding pipelines;

2) 5ml of magnetic beads (2-8 μm in diameter) were added to the sorting vessel 4 using a disposable sterile syringe,

3) Opening a flow control valve 12 (pinch valve) on a first branch pipe 31 connected with a buffer solution bag, a flow control valve 12 on a first second branch pipe 6 and a pump 11, adding buffer solution into a corresponding sorting container 4 from the buffer solution bag, cleaning magnetic beads, removing surface preservative and the like, and then closing the flow control valve 12 and the pump 11;

4) Starting a lifting driving motor 931 to drive the magnet assembly 92 to ascend to the top and adsorb the magnetic beads in the sorting container 4;

5) Opening the flow control valve 12 between the relevant sorting container 4 and one of the collection bags 5 (hereinafter referred to as supernatant bag), opening the rocking drive assembly 82 to rotate the rocking frame 81 to +30 ° for holding, discharging the supernatant in the sorting container 4 into the supernatant bag, and then closing the corresponding flow control valve 12;

6) Opening a flow regulating valve 12 between the cell fluid bag and the pump, the pump 11 and a flow regulating valve 12 on the first second branch pipeline 6, adding 20ml of cell fluid into the sorting container 4 from the cell fluid bag, then closing the flow regulating valve 12 between the cell fluid bag and the pump 11, opening the flow regulating valve 12 between the buffer fluid bag and the pump 11, and adding 180ml of buffer fluid into the sorting container 4 from the buffer fluid bag;

7) The flow regulating valve 12 and the pump 11 between the buffer liquid bag and the pump are closed, the swing driving component 82 is opened to enable the swing frame 81 to swing back and forth from minus 12 degrees to plus 12 degrees, the magnetic beads and the T cells are combined at the speed of about 5 times/min (can be adjusted according to the cell type), the incubation environment temperature is 4 ℃ to 25 ℃, the incubation time is 20-30min,

8) After the incubation time is up, starting a lifting driving motor 931 to drive the magnet assembly 92 to ascend, adsorbing the combination of the magnetic beads and the T cells in the sorting container 4, keeping the cell sap swing, and fully adsorbing for 1min;

9) Keeping the magnetic field open, opening the flow control valve 12 between the sorting container 4 and the supernatant bag, opening the swing driving assembly 82 to rotate the swing frame 81 to +30 degrees for keeping, discharging the supernatant into the supernatant bag, and then closing the flow control valve 12 between the sorting container 4 and the supernatant bag;

10 Opening a flow regulating valve 12 and a pump 11 between the buffer solution bag and the pump, and adding 200ml of buffer solution into the sorting container 4 from the buffer solution bag;

11 Closing the flow control valve 12 and the pump 11 between the buffer bag and the pump, opening the swing driving component 82 to swing the swing frame 81 from-12 DEG to +12 DEG, the speed is about 5 times/min (can also be adjusted according to the cell type), swinging and washing for 1min,

12 Open the flow control valve 12 between the sorting container 4 and the supernatant bag, open the swing driving assembly 82 to rotate the swing frame 81 to +30 ° for holding, discharge the supernatant in the sorting container 4 into the supernatant bag, and then close the flow control valve 12 between the sorting container 4 and the supernatant bag;

13 Open the flow control valve 12 and the pump 11 between the buffer solution bag and the pump, and add 50ml buffer solution into the sorting container 4 from the buffer solution bag;

14 Closing the flow regulating valve 12 and the pump 11 between the buffer bag and the pump, and starting the swing driving assembly 82 to swing the swing frame 81 from-12 DEG to +12 DEG back and forth at a speed of about 5 times/min (which can also be adjusted according to the cell type), so that the combination of the magnetic beads and the T cells is fully swung, vibrated, resuspended and suspended for 1min;

15 The lifting driving motor 931 drives the magnet assembly 92 to descend to the bottom, removes the magnetic field, opens the flow control valve 12 between the sorting container 4 and another collection bag 5 (hereinafter referred to as a cell collection bag), opens the swing driving assembly 82 to rotate the swing frame 81 to +30 ° for holding, discharges the combination of the magnetic beads and the T cells to the target cell collection bag, and then closes the flow control valve 12 between the sorting container 4 and the cell collection bag;

16 ) the sorting process is complete, the system is shut down and the disposable tubing and disposable sorting vessel 4 are disposed of.

Example four:

another cell sorting method is performed by using the cell sorting device of the second embodiment, the two liquid containing bags respectively contain cell liquid and buffer liquid after amplification culture of cell and magnetic bead conjugates, and the sorting container is used for removing magnetic beads in the cell liquid after amplification culture, and the cell sorting method includes the following steps:

y1, adding cell sap: opening the corresponding flow regulating valve 12 to add cell fluid and optionally buffer solution to one or both of the sorting containers 4;

y2, swing of the sorting container 4: the swing mechanism 8 swings the sorting container 4 for a predetermined time to uniformly mix the cells after the amplification culture in the sorting container 4;

y3, discharging a required cell sap target: the magnetic attraction mechanism 9 attracts the magnetic beads or cells and magnetic bead combination objects in the sorting container 4; the swing mechanism 8 tilts the sorting container 4 to a predetermined angle; opening the corresponding flow regulating valve 12 to discharge the required cell sap in the sorting container 4 into a collecting bag 5;

and Y4, repeating Y1 to Y3 for a plurality of times until the cell sap in the liquid containing bag 3 is processed.

Removing magnetic beads after T cell expansion culture:

1) Suspending the liquid containing bag 3 (hereinafter referred to as T cell liquid containing bag) of the T cells after the expanded culture and the liquid containing bag 3 (hereinafter referred to as buffer bag) filled with buffer solution on a hanging frame 13 at the top of the swing frame 81, and connecting the hanging frames with corresponding pipelines;

2) Opening a flow regulating valve 12 and a pump 11 between the T cell liquid containing bag and the sorting container 4, adding 200ml of cell liquid into each of the two sorting containers 4 from the T cell collecting bag after the expanded culture, and then closing the pump 11 to stop injecting;

3) Starting a lifting drive motor 931 to drive the magnet assembly 92 to ascend to the top, and keeping and adsorbing the magnetic beads in the two sorting containers 4;

4) The swing driving assembly 82 is turned on to swing the swing frame 81 back and forth from-12 to +12, the speed is maintained at about 5 times/min (which can also be adjusted according to the cell type),

5) Opening the flow regulating valve 12 between the sorting container 4 and the collection bag 5, opening the swing driving assembly 82 to rotate the swing frame 81 to +30 degrees for keeping, discharging the supernatant to the collection bag 5, and then closing the flow regulating valve 12 between the sorting container 4 and the collection bag 5;

6) Starting the pump 11 again, adding 200ml of cell sap into the sorting container 4 from the T cell collecting bag after the expanded culture, and then closing the pump 11 to stop injecting;

7) The swing driving component 82 is turned on to swing the swing frame 81 back and forth from-12 DEG to +12 DEG, the speed is kept at about 5 times/min (can be adjusted according to the cell type),

8) Opening the flow control valve 12 between the sorting container 4 and the collection bag 5, opening the swing driving assembly 82 to rotate the swing frame 81 to +30 degrees for keeping, discharging the supernatant to the collection bag 5, and then closing the flow control valve 12 between the sorting container 4 and the collection bag 5;

9) Repeating the

steps

6, 7 and 8 until the solution in the T cell collecting bag after the expanded culture is processed,

10 Closing a flow regulating valve 12 between the T cell collecting bag and the pump 11, opening the flow regulating valve 12 between the buffer bag and the pump 11, and the flow regulating valves 12 between the pump 11 and the two sorting containers 4, and adding 50ml of buffer solution into each sorting container 4 from the buffer bag;

11 The flow control valve 12 between the buffer liquid bag and the pump 11 and the flow control valve 12 between the pump 11 and the two sorting containers 4 are closed, the swing driving component 82 is opened to enable the swing frame 81 to swing back and forth from-12 degrees to +12 degrees at a speed of about 5 times/min (can also be adjusted according to the cell type), so that the T cells are fully swung, vibrated, resuspended and the time is 1min;

12 Open the flow control valve 12 between the sorting container 4 and the collection bag 5, open the swing driving assembly 82 to rotate the swing frame 81 to +30 ° for holding, discharge T cells to the target cell collection bag, and then close the flow control valve 12 between the sorting container 4 and the collection bag 5;

the sorting process is complete and the system is shut down and the disposable tubing and disposable sorting vessel 4 are disposed of.

Although the present invention has been described with reference to the preferred embodiments, it is not intended to be limited thereto. The technical solution of the present invention can be used by anyone skilled in the art to make many possible variations and modifications, or to modify equivalent embodiments, without departing from the scope of the technical solution of the present invention, using the technical content disclosed above. Therefore, any simple modification, equivalent change and modification made to the above embodiments by the technical entity of the present invention should fall within the protection scope of the technical solution of the present invention.

Claims

1. A cell sorting device characterized in that: including control box (1), wabbler mechanism (8), be arranged in carrying pump (11) in sorting container (4) with cell liquid, wabbler mechanism (8) are including rocking frame (81) that are used for placing sorting container (4) and swing drive assembly (82) that are used for driving rocking frame (81) to sway, sorting container (4) are equipped with arc portion (43) that are used for bearing cell liquid, be equipped with magnetism on rocking frame (81) and inhale mechanism (9) that are used for changing sorting container (4) magnetic field intensity, be equipped with arc wall (921) with sorting container's arc portion (43) looks adaptation on the magnetism mechanism (9).

2. The cell sorting device according to claim 1, wherein: be equipped with draw-in groove (811) with separation container (4) looks adaptation on wabbler mechanism (8), mechanism (9) are inhaled including locating lift seat (91) on rocking frame (81), locating magnet subassembly (92) on lift seat (91) and being used for driving lift drive assembly (93) that lift seat (91) go up and down, magnet subassembly (92) are located the below of draw-in groove (811), arc wall (921) is formed in the upper surface of magnet subassembly (92).

3. The cell sorting apparatus according to claim 2, wherein: the lifting driving assembly (93) comprises a lifting driving motor (931), a gear (932) and a rack (933), the lifting driving motor (931) is installed on the rocking frame (81), the gear (932) is connected with an output shaft of the lifting driving motor (931), and the rack (933) is arranged on the lifting seat (91) and meshed with the gear (932).

4. The cell sorting apparatus according to claim 3, wherein: be equipped with guide way (812) on rocking frame (81), be equipped with rolling member (911) on lift seat (91), rolling member (911) and the inner wall contact of guide way (812), the both ends of guide way (812) are equipped with locating part (813) respectively.

5. The cell sorting apparatus according to claim 1, wherein: the control box (1) is provided with a digital display module (15) which is electrically connected with each other and a code scanning gun (16) which is used for scanning the code liquid containing bag (3) and the collecting bag (5).

6. The cell sorting device according to any one of claims 2 to 5, wherein: two clamping grooves (811) are formed in the swing frame (81), and the two clamping grooves (811) are arranged in parallel.

7. The cell sorting apparatus according to claim 6, wherein: cell sorting unit still includes trunk line (2), two first minute pipeline (31), two flourishing liquid bag (3), two sorting containers (4), two collection bag (5), two second minute pipeline (6), two third minute pipeline (7) and two fourth minute pipeline (51), flourishing liquid bag (3) are hung and are established on stores pylon (13), collect bag (5) and place on placing platform (14), two in sorting container (4) block respectively locates two draw-in grooves (811) on rocking frame (81), two flourishing liquid bag (3) are connected with the entrance point of pump (11) through two first minute pipeline (31) respectively, two the inlet end of sorting container (4) is connected with the other end of the exit end of pump (11) through two second minute pipeline (6) respectively, goes out the liquid end and is connected with the one end of trunk line (2) through two third minute pipeline (7) respectively, two collection bag (5) are connected with the other end of trunk line (2) through two fourth minute pipeline (51) respectively, each minute pipeline is gone up and all is equipped with governing valve (12 flow).

8. The cell sorting apparatus according to claim 7, wherein: the sorting container (4) is provided with a magnetic bead injection port (41) and a breathing ventilation port (42).